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L00509 Anoplorisk 2014 .pdf

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Title: Risk Management for the EC listed Anoplophora species, A. chinensis and A. glabripennis. (ANOPLORISK)
Author: Phil Northing

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Risk Management for the EC listed Anoplophora species, A.
chinensis and A. glabripennis. (ANOPLORISK)


Final Draft Report



Executive Summary
WP1: Project Management and Co-ordination
Alongside the management of this project, the key role was to encourage interaction
between the partners within the project and between the project partners and other projects
and organisations that have an interest and involvement in understanding Anoplophora spp.
There were three project meetings during the lifetime of this project:
1. Kick off meeting at Fera, York, UK on the 31st January and the 1st February 2011
2. Mid-project meeting at CRA-ABP, Florence, Italy on the 14th and 15th of December
3. Final meeting at JKI, Braunsweig, Germany on the 4th and 5th of December 2012
Two visits to significant outbreak sites were organised to help members of the consortium
experience and understand the impact of long term outbreaks in different situations.
1. Worcester, Massachusetts, 22-25 march 2011. ALB outbreak reported in 2008 but
had likely been there since the mid-late 1990’s.
2. Lombardy and Veneto, Italy 12 - 13 Oct 2011. CLB and ALB outbreaks respectively.
Also, the Austrian team and their Anoplophora dogs visited outbreak sites in Austria, Croatia,
Germany, Switzerland and the UK.
These visits to the outbreak sites were of enormous value in helping consortium members
understand the impact and scale of the task facing those with responsibility for dealing with
the attempted management and eradication of these organisms. This was particularly
relevant for the UK visitors to these sites as they were able to implement some of the lessons
learned on the visits to the outbreak at Paddock Wood in Kent, UK which was found during
the lifetime of this project. The response and subsequent probable eradication of the
outbreak was influenced by knowledge gained during these visits and by other aspects of the
projects work.

WP2: Development and testing of non-destructive detection methods
There are three main topics investigated:

Assessment of different image guided methods for their ability to detect an
Anoplophora infestation in young host plants and in cut wood (P2 and P6)


Assessment of acoustic based detection system to detect and record noises made by
feeding of xylophagous larvae in wood. (P1 with contribution of P5 and P9)
Use of special trained dogs for scenting Anoplophora (ABL/CLB) stages in infested
plants and wood (P3)

The import requirements of the European Union concerning host plants of Anoplophora
chinensis (CLB) include as one of the inspection methods the obligatory destruction of a
specified number of plants of each consignment. Destructive sampling of plants became
necessary because visual inspections led to false negative decisions on several occasions
and therefore infested plants were released. However, destructive sampling is not ideal as it
is costly to the importer and time consuming for the inspector. Effective non-destructive
techniques could ensure that the inspection process remains as effective but provides
significant efficiencies to both the inspectors and the importers. In addition visual inspection
of mature trees in demarcated areas sometimes is difficult and infested trees have been
missed during human based inspection. Aim of the current work package was to assess a
range of new potential techniques which are already in use for other purpose e.g. x-rays,
computed tomography, ultrasound and to further develop a range of promising techniques
that are already partially developed e.g. acoustics and detection dogs.
Work of P1: The work undertaken by Fera and the University of York has further developed
the acoustic detection system which has been tested in various situations from laboratory
(York and FERA) through tests on native trees to use on imported Bonsai and recordings
taken in Italy. Various sensors have been designed and tested with different resonant
frequencies, housing design (including custom housings) and amplification factors. 8-channel
and 16-channel multiplexed systems were developed for use by PSHI on imported Bonsai
and a protocol for their use has been drawn up. The systems have been found to be
successful and have additionally been employed in the laboratory at FERA for long term
recording. Testing of the bite detection software has highlighted that there is still more work
required to make this an effective, efficient system. A sound library for feeding sounds has
been created for 11 species of wood boring beetle including Anoplophora glabripennis, A.
chinensis, Agrilus planipennis, several bark beetles and one Lepidopteran larva (C. cossus).
Discrimination between species has been shown to be feasible during a previous research
project, but the software to enable this is no longer available. Discovery of the presence of
Otiorhynchus sulcatus larvae in the plants at FERA initially suggested that their feeding
sounds are very similar to ALB; subsequent analysis has shown that this is sometimes the
case highlighting the need for discrimination software if all the benefits of an acoustic
detection approach are to be maximised. Two types of artificial larva have been designed
and tested for use with the sensing systems. It is possible for a two sensor system to locate
the approximate position of a larva to within 8-9cm if between the sensors, or whether it is on
the distal side of either sensor. Higher resolution can only be attained if a much higher
sampling rate than 44.1kHz is used. A forward-looking analysis of new systems and
applications has been carried out indicating that future systems can be in two forms – standalone and wireless networks, Each system has application in different but overlapping
Work of P 2: Research performed by NVWA & associate partners - Wageningen University
and Philips Research - was based on analysis of X-ray images recorded on a system
dedicated to luggage inspection (2D) and human health care (3D) respectively. Results show
an increase in accuracy of inspection potential using X-ray instruments. Using the images of

a 2D luggage inspection scanner optimization of the automated borehole detection method
resulted in an accuracy of 67%, Combining machine vision and human input resulted in a
further increase in accuracy up to 83%. Scanning infested wood with a (high-end) Philips
Brilliance iCT scanner for 3D imaging showed (Mol & Wolf, 2011) that: 1) air-filled bore holes
down to 2mm in wooden samples can easily be visualized by CT imaging, 2) the shapes and
sizes of bore holes in infested wood can be visualized in various 3D-display modes, 3)
(dried) larvae of long horned beetles can be identified and visualized, 4) pseudo Anoplophora
bore holes in large (40 unit) bunches of small trees can be spotted visually in the CT crosssectional images. Successful application of 2D and CT scanners for the inspection task at
hand seems technically feasible, but current scanners are not yet for feasible for regular
phytosanitary inspections. To cover any future implementation, it will be necessary to identify
further (inspection) applications for CT imaging and to optimize equipment for the
phytosanitary environment.
Work of P 3: The detection of the scent of A. glabripennis and A. chinensis by detection dogs
is independent from the development stage and the activity of the pest. The four Austrian
Anoplophora detection dogs were successfully used for investigation of wood packaging
material in ports and at stone importers, for the checking of imported plants, also Bonsai, in
nurseries, garden centers and importers as well as for monitoring in infestation areas of A.
glabripennis and A. chinensis in the European states Austria, Netherlands, Italy, Croatia,
Switzerland, Germany and United Kingdom. A training program was developed and already
13 additional dogs from Austria, Germany and Switzerland were educated for the detection
of Anoplophora.
Work of P 6: A CT-scanner developed for assessment of wood properties, several infrared
thermography cameras, radar, electrical resistivity tomography and ultrasonic were
investigated on their potential to detect insect stages as well as boreholes in young plants.
For the trails a model system for host/insect was developed using standardized small logs
cut from young trees and the goat moth (Cossus cossus) larvae. Each method has been
tested on its ability to detect bore holes (with and without larvae) of different diameter and
location in the stem. Infrared thermography could not detect neither larvae nor boreholes.
Radar is dependent on the movement of the test objects; therefore boreholes could not be
detected, but larvae motion even with short mandible movement was detectable. Electrical
resistivity tomography showed some promising results but needs further development.
Boreholes of small sizes (<5 mm) limited the accuracy of this method and it was not possible
in all cases to distinguish larvae from surrounding wood. With the used ultrasonic equipment
borehole size also limited the accuracy of the method. As already stated by P2 above the CT
scanner gave the best results and ended up in a100% accuracy for borehole detection down
to 2 mm, larval identification as well as the identification of frass.
In conclusion X-Rays and CT-Scanner have a high potential to be used for routine inspection
of young plants. The rays (absorbed energy) do not harm the plants and the technology is
independent of the physiological stage of the organisms in the trees. Acoustic detection is
also far developed. Its application depends on the ability of the software to exclude ambient
noise and on the activity of the insect. Detection dogs showed their potential to be used to
detect ALB and CLB as well and became already a routine method either for inspection of
import consignments (plants and wood packaging) or for analysis of infested trees in the wild.
Within this WP, the following papers were published:


1. Hoffmann, N.; Schröder, T. (2012): Potential of infrared thermography to detect insect
stages and defects in young trees. Julius-Kühn-Archiv, 438: 166-167.
2. Hoffmann, N; Schröder, T. (in press): Potential of infrared thermography to detect insect
stages and defects in young trees. Journal für Kulturpflanzen.
3. Hoyer-Tomiczek, U (2011).: Früh erkennen und wenig zerstören: Spürhunde erschnüffeln
Baumschädlinge. Risikomanagement für die in der EG gelisteten Anoplophora-Arten
2011 (ANOPLORISK). Jahresbericht 2011, Bundesforschungs- und Ausbildungszentrum
für Wald, Naturgefahren und Landschaft (BFW), Wien, S.27.
4. Hoyer-Tomiczek, U., Sauseng, G.: 2012. "Alternative Detection Methode for ALB and
CLB". 2012. Fortschutz Aktuell 55, Bundesforschungs- & Ausbildungszentrum für Wald,
Naturgefahren und Landschaft (BFW), Wien, 2012, S 43-45.
5. Hoyer-Tomiczek, U., Krehan, H., Hoch, G.: (in press). Wood boring insects in wood
packing material: Recent interceptions and a new ALB outbreak in Austria. Abstract
USDA, 2013.
6. Jansen, R.M.C. & Hemming, J. (2011). “Validation of X-ray for borehole detection in
intact trees” Draft Wageningen UR Greenhouse Horticulture, Wageningen / Bleiswijk,
December 2011, 26 pp.
7. Mol, C.R. & Wolf, R.M. (2011) “3D imaging for the detection of Anoplophora in wood”
Report Philips Research, Eindhoven, August 2011, 30pp.

WP3: Development and testing of diagnostic techniques in the absence of the
There were five main topics investigated
1. Dissection of symptomatic wood in the laboratory to obtain insect frass and investigation
of the collected frass for the presence of body parts of insects using stereomicroscope
2. Diagnosis/confirmation of Anoplophora spp. based on molecular analysis of insect body
3. Analysis of other Anoplophora species
4. Non destructive diagnosis of Anoplophora spp. tree colonization based on analysis of
insect frass
5. Dissection of symptomatic plants and analysis of the annual ring in laboratory to date exit
The first two topics were performed by P7 (ILVO) and permit to define a correct way to
extract, from the wooden parts of plants infested by woodboring insects, remnants of the
body of the insect (head capsule and larval skins), using tools and methods to split the wood
and by the use of laboratory instruments to detect and sample the insect remnants
(stereomicroscopes and the most appropriate magnification, forceps and containers). On
these insect remnants, it was defined an appropriate molecular analysis protocol for species
Regarding the third topic, the partner P3 (BFW) performed with success molecular analysis
on insects belonging to the genus Anoplophora, but different from A. chinensis and A.
glabripennis. Thanks the possibility to obtain several specimens of exotic species of the
genus Anoplophora from their native oriental region, molecular analysis was performed, and
for the following Anoplophora species species-specific finger prints are now available:

Anoplophora chinensis, Anoplophora chinensis form malasiaca, Anoplophora glabripennis,
Anoplophora beryllina, Anoplophora davidis, Anoplophora elegans, Anoplophora granata,
Anoplophora macularia, Anoplophora sollii.
The fourth topic was performed by P5 (CRA-ABP) and permit to define a correct protocol for
molecular analysis for species identification of woodboring insects (potentially applicable to a
wide range of phytophagous insects) starting from the frass produced by the larvae during
the feeding activity. This procedure poses interesting perspectives for pest identification in
the absence of the insect (absence of adults, larvae or pupae for different reasons, or when
in a first step, the plant destruction presents several impediments); this method is
presumably applicable for many time after insect emergence from the infested plant, i.e. the
frass contained in the beetle galleries into the wood of the infested plant can be suitable for
molecular analysis for a long period of time (in appropriate environmental conditions), also
for years.
The fifth topic was performed by P5 (CRA-ABP) and investigated a method for dating exit
holes produced by the adults of Anoplophora during their emergence from the infested
plants. The stepwise procedure described permit a correct sample processing, and analysis
procedure necessary to dating the time of exit hole excavation, taking into account that trees
must be alive at time of felling. In this way it is possible to count backwards from the exit
hole, the number of annual growth rings and to define the timing of beetle occurrence. These
information can be useful in infestation dynamics studies.
An additional topic was performed by P5 (CRA-ABP) which was not included originally in the
topics proposed in the project. Within this WP, it was investigated the possibility to prepare a
taxonomic key based on larval morphology aimed to identify, or at least to separate the two
exotic species of the genus Anoplophora present currently in Europe (together with the
recent exotic species Psachotea hilaris), from the other woodboring longhorned beetle of the
native European fauna. This taxonomic key is provided with many detailed morphological
pictures, which aid in the interpretation of the characters mentioned in the key. This key can
be a rapid and useful tool in species identification during phytosanitary surveys (taking into
account that most cited details can be observed in the field using a pocket magnifying glass),
before eventually proceed with molecular analysis in doubtful cases.

Within this WP, the following papers were published:
1) Strangi A., Sabbatini Peverieri G., Rovrsi P.F., (2012). Managing outbreaks of the citrus
long-horned beetle Anoplophora chinensis (Forster) in Europe: Molecular diagnosis of
plant infestation. Pest Management Science, DOI 10.1002/ps.3416.
2) Sabbatini Peverieri G., Bertini G., Furlan P., Cortini G., Roversi P.F., 2012. Anoplophora
chinensis (Forster) (Coleoptera Cerambycidae) in the outbreak site in Rome (Italy):
experiences in dating exit holes. REDIA, XCV: 89-92.
3) Pennacchio F., Sabbatini Peverieri G., Jucker C., Allegro G., Roversi P.F., 2012. A key
for the identification of larvae of Anoplophora chinensis, Anoplophora glabripennis and
Psacothea hilaris (Coleoptera Cerambycidae Lamiinae) in Europe. REDIA XCV: 57-65.



WP4: Understanding the potential for dispersal at outbreak sites
There were 4 main topics investigated:
1. Investigate if A. glabripennis is able to complete its development on fruit trees in Europe
2. Establish whether conifers are suitable host trees for A. chinensis
3. Consider the impact of host density and other environmental variables on the dispersal
and potential spread of Anoplophora species
4. Determine the susceptibility of the most important Citrus spp. to CLB

Monitoring activities in European areas with Anoplophora-infestation and specific host plantstests with both Anoplophora species revealed interesting new results about additional new
host plants.
In Austria the deciduous tree species Fagus sp., Fraxinus sp. and Alnus sp. are new hosts of
A. glabripennis which had not been known as host plants so far in Europe and partially also
in the rest of the world. Alder (Alnus sp.) could also be an additional host where only
oviposition scars and young larvae of A. glabripennis have been observed but no further
development of them due to cutting of these trees (further development of these young
larvae in the cut branches failed under quarantine laboratory conditions of BFW). An ALBinfestation of fruit trees has never been observed during a monitoring period of 11 years in
Upper Austria with exception of one finding in a new infestation area in 2012 where one
Prunus avium tree showed oviposition scars and starting larval galleries, but no larvae.
Specific testing of apple trees (Malus domestica, cultivar: Golden Delicious) showed that this
fruit tree is a suitable host-plant for complete development of Anoplophora glabripennis
(ALB) under controlled conditions, but it is not proved for natural field conditions. The
complete development of ALB is even possible in trees with stems of only 3 cm diameter.
There is a potential risk in apple production (Golden Delicious) in Europe in case of ALBinfestation - especially if there is a „lack“ of „better“ hosts.
The Italian testing of conifers and deciduous trees including Citrus species as suitable hosts
for A. chinensis (CLB) showed clearly its polyphagy and adaptation to many tree species. In
fact, it was possible to observe within the field trial that adults fed on all tested Citrus species
(C. x sinensis, C. reticulata and C. x limon), on Populus x euramericana and Acer
saccharinum and also on conifers (Taxus baccata, Cryptomeria japonica aritaki, but not on
Pinus sylvestris), oviposition scars only were not present on Citrus x limon and larval activity
was present on 5 of 7 tested species with signs of oviposition (Populus x euramericana,
Cryptomeria japonica aritaki, Citrus x sinensis, Citrus reticulata and Acer saccharinum). Due
to the start of the trials in 2012 it is only possible to verify the full development of CLB on
these host plants in the next future, maybe in summer 2013 or 2014.
It can be concluded that deliverable D4.1 (Information about additional unknown host
plants of A. glabripennis and A. chinensis in Europe, especially if fruit trees are
suitable hosts for ALB and coniferous trees for CLB) was fulfilled during this project.
Concerning the achievement of deliverable D4.2 (Review of the current knowledge on the
dispersal behaviour of Anoplophora species; modelling the dispersal capacity of

Anoplophora species in relation to biotic and abiotic conditions at an outbreak site) it
can be stated that it was partially fulfilled.
Preliminary results of an Italian study which was based on data of an infestation area in NEItaly showed that it is unlikely that ALB will spread much farther than 2000 meters according
with previously reported dispersal distance (Smith et al., 2001; 2004).
The Dutch team investigated the influence on dispersal of Anoplophora species by climatic
conditions and estimated their lifecycles using an accumulated Degree Day Model. Thus, the
development of Anoplophora beetles under Dutch conditions will take on average 3 years.
Main Conclusions
The obtained results and knowledge about new suitable host plants of Anoplophora species
should be of high importance for monitoring and eradication measures in every infestation
area in Europe and have to be considered when taking decisions on the surveys or
eradication programs.
Papers, other publications and dissemination activities done
Oral presentation about preliminary results at the Austrian Plant Protection meeting
(Österreichische Pflanzenschutztage 2011) in St. Pölten, 30.11.2011:
Lethmayer, C. & Hoyer-Tomiczek, U.: Asiatischer Laubholzbockkäfer (Anoplophora
glabripennis) – auch eine Gefahr für den Apfelanbau in Österreich?
(= Asian long-horn beetle (Anoplophora glabripennis) – also a danger for the Austrian apple

WP5: Investigating the potential efficacy of different management practices
There were 3 main topics for investigation:
1. Review the available information on management practices and attempt to fill relevant
2. Evaluate the potential of a range of chemical treatment / treatment methods
3. Evaluate the potential for utilisation of biological control and/or enhancement of natural

A desk study was undertaken to collate and interpret the current knowledge on the potential
for controlling/managing infestations of Anoplophora spp (longhorned beetles). A review of
the available information aimed to provide information on current management practices,
identify relevant knowledge gaps, evaluate the potential of a range of chemical / treatment
methods and evaluate the potential for utilisation of biological control.
There is a wealth of information on the variety of techniques that have been investigated to
prevent, control and eradicate Anoplophora infestations, including cultural (e.g. siviculture,
tree management, pest resistant clones), chemical (e.g. systemic, contact, fumigation),
physical (e.g. heat, irradiation, exclusion) and biological (e.g. fungi, nematodes, bacteria,
parasitoids) measures. Those that have received the greatest attention and which show the
greatest potential for use in a practical situation, have been considered in this review.

Eradication is the goal for all infestations, however an integrated approach is essential for
that goal to be achieved. The main strategies that have been vital to achieving eradication of
Anoplophora infestations include tree removal and chipping, the use of protective
insecticides, and public involvement.
Further research should investigate maximising the efficacy of existing control measures and
investigating some of the more novel control measures, such as effects on the beetles’
metabolism. The use of fungal bands may offer the most promising use of biological control
agents, with efficacy enhanced by combining with an attractant or pheromone. The potential
exists to augment eradication efforts by combining different methodologies. Other areas to
consider may include pesticide rotation to reduce the risk of pesticide resistance and the use
of alternatives methods for the treatment of wood packaging.
The longhorned beetle species Anoplophora chinensis and Anoplophora glabripennis have
emerged in the last two decades as a risk to urban and woodland trees in Europe and there
have been several outbreaks of these beetles in Europe. This review summarises the
literature on biocontrol of Anoplophora spp. and discusses which are the strongest
candidates for use in Europe. Some of the methods below could be useful for control, but are
unlikely to be instrumental in achieving eradication. Below is a summary of the findings:
Entomopathogenic fungi: Strong candidate as a biopesticide as fungal infection results
in high mortality rates and has already been developed into a commercial product in
Japan for Anoplophora control. Beauvaria bassiana, is already authorised for use as a
biopesticide in the UK.
Parasitic nematodes: Another strong candidate for use as a biopesticide due to high
mortality rates and effective application methods have already been developed. Two
nematode species, S. feltiae and S. carpocapsae, are also au for use in the UK and other
parts of the EU.
Parasitoids: Several parasitoid species (e.g. Dastarcus helophoroides) have been
shown to be effective biological control agents in China but their specificity would need to
be investigated further before use in Europe. Some native European parasitoid species
(e.g. Spathius erythrocephalus) have potential to be used as a biocontrol.
Predators: Two woodpecker species (Dendrocopos major and Picus canus) have been
shown to be effective at controlling A. glabripennis numbers in Chinese forests and are
also native to Europe.
Pathogenic bacteria: Are currently not a strong prospect for use in biocontrol with no
study yet advancing to field trials.
It was not possible within the scope of this project to undertake any significant R&D to
attempt to fill any gaps in available information.

WP6: Investigating the biology of Anoplophora spp. in relevant EU climatic
There were 5 main topics for investigation
1. Review current knowledge on the link between climate and the potential for Anoplophora
population development.

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